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一种用于快速同时检测临床标本中的猪捷申病毒、猪瘟病毒和猪繁殖与呼吸综合征病毒的多重 RT-PCR 方法。

A multiplex RT-PCR for rapid and simultaneous detection of porcine teschovirus, classical swine fever virus, and porcine reproductive and respiratory syndrome virus in clinical specimens.

机构信息

Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, No. 427, Maduan Street, Nangang District, Harbin 150001, China.

出版信息

J Virol Methods. 2011 Mar;172(1-2):88-92. doi: 10.1016/j.jviromet.2010.12.023. Epub 2010 Dec 28.

Abstract

A multiplex RT-PCR (mRT-PCR) assay was developed and evaluated for its ability to detect multiple viral infections of swine simultaneously. One pair of primers was selected carefully for each of the following three RNA viruses: porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), and porcine teschovirus (PTV). Each target produced a specific amplicon with a size of 451bp (PRRSV), 343bp (CSFV), or 163bp (PTV). The sensitivity of the mRT-PCR using purified plasmid constructs containing the specific viral target fragments was 2.02 x 10², 2.90 x 10³, and 6.16 x 10³ copies for PRRSV, CSFV, and PTV, respectively. Among 69 clinical samples from Heilongjiang, Jilin, and Henan provinces, co-infection by PRRSV and CSFV was 4.4%, co-infection by PRRSV and PTV was 11.6%, co-infection by PTV and CSFV was 13.0%, and co-infection by the three viruses was 8.7%. In conclusion, the mRT-PCR should be useful for routine molecular diagnosis and epidemiology.

摘要

建立了一种多重 RT-PCR(mRT-PCR)检测方法,用于同时检测猪的多种病毒感染。针对以下三种 RNA 病毒:猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)和猪传染性胃肠炎病毒(PTV),分别精心选择了一对引物。每个靶标都产生了一个特定的扩增子,大小分别为 451bp(PRRSV)、343bp(CSFV)或 163bp(PTV)。使用包含特定病毒靶片段的纯化质粒构建体进行 mRT-PCR 的灵敏度分别为 PRRSV 为 2.02 x 10²、CSFV 为 2.90 x 10³和 PTV 为 6.16 x 10³拷贝。在来自黑龙江、吉林和河南三省的 69 份临床样本中,PRRSV 和 CSFV 的混合感染率为 4.4%,PRRSV 和 PTV 的混合感染率为 11.6%,PTV 和 CSFV 的混合感染率为 13.0%,三种病毒的混合感染率为 8.7%。总之,mRT-PCR 应该有助于常规分子诊断和流行病学。

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